Screening assay

Abstract

The present invention relates to agents that modulate the interaction of a dopamine receptor interacting polypeptide (DRIP) as represented by FIGS. 1 to 11 and to screening methods for the identification of such agents.

1 . A screening method for the identification of agents that modulate the interaction of a polypeptide with one or more dopamine receptors wherein said polypeptide is selected from the group consisting of: i) a polypeptide, or fragment or variant thereof, encoded by a nucleic acid molecule consisting of a nucleic acid sequence as represented by  FIG. 1 ,  2 ,  3 ,  4 ,  5 ,  6 ,  7 ,  8 ,  9   10  or  11 , or a sequence complementary thereto, or a fragment thereof; ii) a polypeptide encoded by a nucleic acid molecule which hybridises to a nucleic acid molecule as defined in (i) above and which has dopamine receptor binding activity; and iii) a polypeptide comprising a nucleic acid which is degenerate as a result of the genetic code to the nucleic acid sequence defined in (i) and (ii); comprising the steps of i) forming a preparation comprising said polypeptide and a dopamine receptor; and ii) adding at least one candidate agent to be tested; iii) determining the effect, or not, or said agent on the interaction of said polypeptide with said dopamine receptor. 
     
     
         2 . A method as claimed in  claim 1  wherein the polypeptide is represented by the amino acid sequence as shown in  FIG. 1 ,  2 ,  3 ,  4 ,  5 ,  6 ,  7 ,  8 ,  9   10  or  11 , or a variant thereof. 
     
     
         3 . A method as claimed in  claim 1  wherein the nucleic acid molecule anneals under stringent hybridisation conditions to the nucleic acid sequence shown in  FIG. 1 ,  2 ,  3 ,  4 ,  5 ,  6 ,  7 ,  8 ,  9 ,  10  or  11  or to its complementary strand. 
     
     
         4 . A method as claimed in  claim 1  wherein the nucleic acid sequence comprises nucleotides 644 to 1191 of the sequence shown in  FIG. 1 . 
     
     
         5 . A method as claimed in  claim 1  wherein the nucleic acid sequence comprises nucleotides 1 to 601 of the sequence shown in  FIG. 2 . 
     
     
         6 . A method as claimed in  claim 1  wherein the nucleic acid sequence comprises nucleotides 1 to 521 of the sequence shown in  FIG. 3 . 
     
     
         7 . A method as claimed in  claim 1  wherein the nucleic acid sequence comprises nucleotides 415 to 910 of the sequence shown in  FIG. 4 . 
     
     
         8 . A method as claimed in  claim 1  wherein the nucleic acid sequence comprises nucleotides 1058 to 1661 of the sequence shown in  FIG. 6 . 
     
     
         9 . A method as claimed in  claim 1  wherein the nucleic acid sequence comprises nucleotides 621 to 1069 of the sequence shown in  FIG. 7 . 
     
     
         10 . A method as claimed in  claim 1  wherein the dopamine receptor is represented by a polypeptide having the amino acid sequence shown in  FIG. 12 ,  13 ,  14 ,  15  or  16 . 
     
     
         11 . An agent wherein said agent modulates the interaction of a polypeptide with said dopamine receptor wherein said polypeptide is selected from the group consisting of: i) a polypeptide, or fragment or variant thereof, encoded by a nucleic acid molecule consisting of a nucleic acid sequence as represented by  FIG. 1 ,  2 ,  3 ,  4 ,  5 ,  6 ,  7 ,  8 ,  9   10  or  11 , or a sequence complementary thereto, or a fragment thereof; ii) a polypeptide encoded by a nucleic acid molecule which hybridises to a nucleic acid molecule as defined in (i) above and which has dopamine receptor binding activity; and iii) a polypeptide comprising a nucleic acid which is degenerate as a result of the genetic code to the nucleic acid sequence defined in (i) and (ii). 
     
     
         12 . An agent as claimed in  claim 11  wherein the agent binds a dopamine receptor and wherein the agent is not the natural ligand of a dopamine receptor. 
     
     
         13 . An agent as claimed in  claim 11  wherein said agent is an antibody or an active binding fragment of a monoclonal antibody. 
     
     
         14 . An agent as claimed in  claim 13  wherein the antibody fragment is a single chain antibody variable region fragment. 
     
     
         15 . An agent as claimed in  claim 13  wherein the antibody or fragment binds a polypeptide comprising an amino acid sequence as represented by  FIG. 1 ,  2 ,  3 ,  4 ,  5 ,  6 ,  7 ,  8 ,  9   10  or  11 , or a variant thereof. 
     
     
         16 . An agent as claimed in  claim 13  wherein the antibody fragment binds a polypeptide encoded by nucleotides 644 to 1191 of the sequence shown in  FIG. 1 . 
     
     
         17 . An agent as claimed in  claim 13  wherein the antibody fragment binds a polypeptide encoded by nucleotides 1 to 601 of the sequence shown in  FIG. 2 . 
     
     
         18 . An agent as claimed in  claim 13  wherein the antibody fragment binds a polypeptide encoded by nucleotides 1 to 521 of the sequence shown in  FIG. 3 . 
     
     
         19 . An agent as claimed in  claim 13  wherein the antibody fragment binds a polypeptide encoded by nucleotides 415 to 910 of the sequence shown in  FIG. 4 . 
     
     
         20 . An agent as claimed in  claim 13  wherein the antibody fragment binds a polypeptide encoded by nucleotides 1058 to 1661 of the sequence shown in  FIG. 6 . 
     
     
         21 . An agent as claimed in  claim 13  wherein the antibody fragment binds a polypeptide encoded by nucleotides 621 to 1069 of the sequence shown in  FIG. 7 . 
     
     
         22 . An agent as claimed in  claim 13  wherein the antibody fragment binds a polypeptide encoded by nucleotides 1710 to 2051 of the sequence shown in  FIG. 12 . 
     
     
         23 . An agent as claimed in  claim 13  wherein the antibody fragment binds a polypeptide encoded by nucleotides 1356 to 1523 of the sequence shown in  FIG. 12 . 
     
     
         24 . An agent as claimed in  claim 13  wherein the antibody fragment binds a polypeptide encoded by nucleotides 796 to 1281 of the sequence shown in  FIG. 13 . 
     
     
         25 . An agent as claimed in  claim 13  wherein the antibody fragment binds a polypeptide encoded by nucleotides 1005 to 1364 of the sequence shown in  FIG. 14 . 
     
     
         26 . An agent as claimed in  claim 13  wherein the antibody fragment binds a polypeptide encoded by nucleotides 646 to 1023 of the sequence shown in  FIG. 15 . 
     
     
         27 . An agent as claimed in  claim 13  wherein the antibody fragment binds a polypeptide encoded by nucleotides 772 to 948 of the sequence shown in  FIG. 16 . 
     
     
         28 . An agent as claimed in  claim 13  wherein the antibody fragment is a chimeric antibody. 
     
     
         29 . An agent as claimed in  claim 13  wherein the antibody fragment is a humanised antibody. 
     
     
         30 . An agent as claimed in  claim 11  wherein the agent is a peptide such as a modified peptide. 
     
     
         31 . An agent as claimed in  claim 30  wherein the peptide has an amino acid sequence encoded by nucleotides 1710 to 2051 of the sequence shown in  FIG. 12 . 
     
     
         32 . An agent as claimed in  claim 30  wherein the peptide has an amino acid sequence encoded by nucleotides 1356 to 1523 of the sequence shown in FIG. 
     
     
         33 . An agent as claimed in  claim 30  wherein the peptide has an amino acid sequence encoded by nucleotides 796 to 1281 of the sequence shown in FIG. 
     
     
         34 . An agent as claimed in  claim 30  wherein the peptide has an amino acid sequence encoded by nucleotides 1005 to 1364 of the sequence shown in FIG. 
     
     
         35 . An agent as claimed in  claim 30  wherein the peptide has an amino acid sequence encoded by nucleotides 646 to 1023 of the sequence shown in FIG. 
     
     
         36 . An agent as claimed in  claim 30  wherein the peptide has an amino acid sequence encoded by nucleotides 772 to 948 of the sequence shown in  FIG. 16 . 
     
     
         37 . An agent as claimed in  claim 11  wherein the agent is an aptamer. 
     
     
         38 . An agent as claimed in  claim 11  wherein the agent is an interfering RNA (RNAi) molecule. 
     
     
         39 . A cell transfected with at least one nucleic acid molecule wherein the genome of said cell is modified to include at least one copy of a nucleic acid molecule encoding a polypeptide selected from the group consisting of i) a polypeptide, or fragment or variant thereof, encoded by a nucleic acid molecule consisting of a nucleic acid sequence as represented by  FIG. 1 ,  2 ,  3 ,  4 ,  5 ,  6 ,  7 ,  8 ,  9   10  or  11  or a sequence complementary thereto, or a fragment thereof; ii) a polypeptide encoded by a nucleic acid molecule which hybridises to a nucleic acid molecule as defined in (i) above and which has dopamine receptor binding activity; iii) a polypeptide comprising a nucleic acid which is degenerate as a result of the genetic code to the nucleic acid sequence defined in (i) and (ii) wherein said cell is adapted for the regulated expression of said nucleic acid molecule. 
     
     
         40 . A cell as claimed in  claim 39  wherein said cell is further transfected with at least one nucleic acid molecule wherein the genome of said cell is modified to include at least one copy of a nucleic acid molecule encoding one or more dopamine receptors. 
     
     
         41 . A cell as claimed in  claim 39  wherein said cell is a brain cell. 
     
     
         42 . A cell as claimed in  claim 39  wherein said cell is a neuronal cell. 
     
     
         43 . A transgenic non-human animal comprising at least one cell as claimed in  claim 39 . 
     
     
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         46 . (canceled) 
     
     
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         48 . (canceled) 
     
     
         49 . (canceled) 
     
     
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         53 . (canceled)